Fig. 2: SGS NPCs display features of SETBP1 accumulation.

a NPC generation protocol (upper panel) and representative bright-field images of isogenic control (D868D) and mutant (D868N) NPCs. b Representative images (taken at the same magnification) of NESTIN (P = 0.3897) and PAX6 (P = 0.8436) immunostaining and relative quantification. n = 3. See also Supplementary Fig. 2a. c SETBP1 immunoblotting in WT, isogenic control (D868D and I871I) and mutant (D868N and I871T) NPCs and relative quantification. * indicates statistic test between isogenic cell line pairs. P = 0.0048; WT vs. D868D P = 0.9686; WT vs. D868N P = 0.2983; WT vs. I871I P = 0.3912; WT vs. I871T P = 0.6119; D868D vs. D868N *P = 0.0465; I871I vs. I871T *P = 0.0361; D868N vs. I871T P = 0.9966. WT vs. MUT ***P = 0.0004. n = 3. d SET immunoblotting in WT, isogenic control and mutant NPCs and relative quantification * indicates statistic test between isogenic cell line pairs. ^# refers to the comparison between each mutant cell line with the unrelated WT. ° refers to the comparison between mutant NPCs. n = 3. See Supplementary Dataset 3 for details of statistical analysis. e r total and phosphorylated PP2A (pPP2A Tyr307) immunoblotting in WT, isogenic control and mutant NPCs and relative quantification of pPP2A/PP2A ratio. P = 0.7713. D868D vs. D868N P = 0.8981, I871I vs. I871T P = 0.9576; See Supplementary Fig. 2e for WT vs. MUT comparisons. n = 5. f Phosphatase activity assay of PP2A in isogenic control and mutant NPCs. P = 0.1343, D868D vs. D868N #1 P = 0.1315, D868D vs. D868N #2 P = 0.2760. #1 and #2 refer to two independent clones of the D868N IPSC line. n = 3. All data expressed as mean ± SEM. ns: nonsignificant differences when P > 0.05; ^∗P < 0.05, ^∗∗P < 0.01, ^∗∗∗P < 0.001, ^∗∗∗∗P < 0.0001. b–f One-way ANOVA followed by Tukey post hoc test for multiple comparisons, except for WT vs. MUT comparisons two-sided unpaired t test in c and d.